Eect of dierent temperatures and storage atmospheres on

Coratina olive oil quality

Maria Lisa Clodoveo
a,
*
, Debora Delcuratolo
a
, Tommaso Gomes
a
, Giancarlo Colelli
b
a
Department of Engineering and Management of the Agricultural, Livestock and Forest Systems, Agriculture Faculty,
University of Bari, Via Amendola 165/a, 70126 Bari, Italy
b
Production Sciences, Engineering and Economics for Agricultural Systems Department, Agriculture Faculty, University of Foggia,
Via Napoli 25, 71100 Foggia, Italy
Received 25 November 2005; received in revised form 12 May 2006; accepted 21 May 2006
Abstract
Olives (Olea europaea cv. Coratina) used for oil production were stored for 30 days at three dierent temperatures and under dierent
atmospheres (ambient temperature, 5 C with a ux of humidied air, 5 C with a ux of 3%O
2
+ 5%CO
2
). The olives were kept in jars
used for fruit storage, each with a capacity for 1.5 kg of olives.
Conventional analyses (acidity, peroxide value, specic extinction coecient at 232 nm and 270 nm) and non conventional (polar
compounds) analyses were carried out in order to measure the hydrolytic and oxidative degradation of the oils obtained from the olives.
Storage at 5 C, both under a ow of humidied air and a ow of 3%O
2
+ 5%CO
2
, produced oils that maintained their initial chemical
qualities until the end of the experimentation; whereas storage of olives at room temperature led to their deterioration after 15 days of
storage.
2006 Elsevier Ltd. All rights reserved.
Keywords: Olea europaea; Post-harvest; Olive oil quality; High-performance size-exclusion chromatography; Oligopolymers; Polar compounds
1. Introduction
Olive oil quality is directly related to the physiological
conditions of the olives from which the oil is extracted.
Olive processing in important producing countries (such
as Spain, Italy, and Greece) is often not well synchronized
with crop harvests (Garcia and Streif, 1991; Gutierrez,
Perdiguero, Garcia, and Castellano, 1992). Olives are often
piled into large heaps and stored at ambient temperature
for up to several weeks prior to processing for oil extrac-
tion (Garcia, Gutierrez, Castellano, Perdiguero, and Albi,
1996), and this is when the greatest deterioration takes
place (Olias and Garcia, 1997). Pressure within the olive
pile during storage may cause secretion of uid from the
olives thereby providing an optimum medium for the
growth of fungi and bacteria (Olias and Garcia, 1997).
Under these conditions, anaerobiosis can occur in the inner
part of the pile while aerobic losses occur in the outer part
(Garcia and Streif, 1991). Furthermore, heat production
from respiratory activity may accelerate the deterioration
of the fruit (Garcia and Streif, 1991) and eventually cause
the breakdown of cell structure (Gutierrez, Perdiguero,
Garcia et al., 1992). Oil extracted from these damaged
olives can be high in acidity and low in stability (Garcia,
Gutierrez, Castellano et al., 1996) and can develop a great
amount of volatile acids (acetic or butyric) that cause a
characteristic musty smell (Olias and Garcia, 1997). In a
few days, the physical and chemical structure of the olives
is altered and the oil extracted from them has a very poor
quality. This type of oil must be rened before consump-
tion. (Gutierrez, Perdiguero, Garcia et al., 1992). A great
number of dierent methods for olive storage have been
experimented to nd ways to counter this important
0308-8146/$ - see front matter 2006 Elsevier Ltd. All rights reserved.
doi:10.1016/j.foodchem.2006.05.035
*
Corresponding author. Tel.: +39 80 544 2941; fax: +39 80 544 3015.
E-mail address: clodoveo@agr.uniba.it (M.L. Clodoveo).
www.elsevier.com/locate/foodchem
Food Chemistry 102 (2007) 571576
Food
Chemistry
problem (Castellano, Garcia, Morilla, Perdiguero, and
Gutierrez, 1993; Garcia, 1993a, 1993b; Garcia, Gutierrez,
Barrera, and Albi, 1996; Garcia et al., 1994; Kader, 1986;
Kader, Nanos, and Kerbel, 1989, 1990; Koprivnjak, Conte,
and Totis, 2002; Pereira, Casal, Bento, and Oliveira, 2002;
Petruccioli and Parlati, 1987).
Very few articles concerning polar compound concen-
trations or HPSEC analyses of virgin oils from stored Pic-
ual olives are available (Perez-Camino, Garcia, &
Castellano, 1992) and overall, little information is found
on the storage of Italian cultivars. It is possible that mill
olive varieties behave dierently under refrigeration. Our
study was conducted to identify the optimum pre-process-
ing storage temperature and atmospheric composition to
preserve the quality of Coratina olives and oil.
2. Materials and methods
2.1. Experimental material
Olive fruits (Olea europaea L.) from the Coratina variety
were harvested in olive groves in the same area near Foggia
(Apulia Italy) during the crop season 2002/2003. The
olives were randomly picked at the industrial optimal rip-
ening stage, according to their skin colour. Harvesting
was done by hand, using rakes. Olives were sorted to
obtain fruits of uniform size and colour and then distrib-
uted randomly in 1.5 kg lots that were placed into 14 glass
jars.
2.2. Storage treatments
A group of six jars of olives were kept at 5 C and con-
nected to a continuous humidied air ow or a
3%O
2
+ 5%CO
2
ow-through-system at a ow rate of
500 ml min
1
. Another group of six jars of olives were kept
at 20 C in normal ambient air (control) to verify the dete-
rioration rate at an ambient temperature. Two replicates
were used per treatment. Each sample was subjected to
three analytical determinations. All data points in the g-
ures are the mean SD. Quality evaluations were made
initially on day 1 and after 15 and 30 days of olive storage.
2.3. Sample preparation
After washing and leaf-removal, each batch was ham-
mer-crushed in a pilot plant. After malaxation for 30 min
at 25 C, the oil was extracted by means of a laboratory
basket centrifuge. All the oil samples were ltered through
cotton and stored at 20 C in the dark in amber glass bot-
tles without head space until analysis.
2.4. Percent free fatty acids, peroxide value and ultra-violet
light absorption
Free fatty acids value, peroxide value and UV light
absorption (K
232
, K
270
) were determined following the o-
cial analytical methods described in EC Regulation 2568/
91 and subsequent modications ECC/796/02 and ECC/
1989/03.
2.5. Total phenol content
Phenolic compounds were isolated from a solution of oil
in hexane by triple-extraction with watermethanol (60:40
v/v). Total phenols, expressed as gallic acid equivalents
(ppm), were determined with a UV visible spectrophotom-
eter (Beckman) at 765 nm using the Folin-Ciocalteu
reagent (Swain and Hillis, 1969).
2.6. Oxidative stability
Oxidation induction time was evaluated by the Ranci-
mat method. Stability was expressed as the oxidation
induction time (h), measured with the Rancimat apparatus
(Metrohm AG, Herison, Switzerland) (T = 120 C; air ow
rate = 20 l/h).
2.7. Polar compounds and triglyceride oligopolymers
Polar compounds (PC) were determined in each sample
by means of silica gel column chromatography as described
by the IUPAC method (IUPAC, 1987). PC were analyzed
by high-performance size exclusion chromatography
(HPSEC) to determine oxidized triglycerides (ox-TG),
oligopolymers, and partial glycerides.
The chromatographic system consisted of a Perkin
Elmer pump, series 10, a 7125 S sample injector
(Rheodyne), a 50 ll injector loop, and a series of three
PL-gel columns (PerkinElmer Ltd., Beaconseld, Great
Britain) of 0.75 cm i.d. 30 cm length. The columns
were packed with highly cross-linked styrene divinylben-
zene copolymers with a particle diameter of 5 lm and
pore diameters of 500, 500, and 100 A

, respectively. A
PL-gel guard column (PerkinElmer Ltd.) of 7.5 mm
i.d. 5 cm length was used. The detector was a dieren-
tial refractometer (Shimadzu RID 6A, Shimadzu Corp.,
Japan) connected to an integrator. The elution solvent
used was CH
2
Cl
2
for HPLC at a ow rate of 1.0 ml/
min.
The procedures for identifying the peaks on each chro-
matogram and for the quantitative assessment of the clas-
ses of compounds under investigation were carried out as
described elsewhere (Gomes, 1992; Gomes and Caponio,
1999).
2.8. Statistical analysis
Statistical analysis was performed using Microsoft Excel
software. Signicant dierences between treatments were
determined using one-way ANOVA followed by Dun-
cans test (p < 0.05) carried out on the Software Stat-
graphics Plus (Manugistics, MD, USA).
572 M.L. Clodoveo et al. / Food Chemistry 102 (2007) 571576
3. Results and discussion
3.1. Percent free fatty acids
Virgin olive oil contains about 98% neutral lipids,
mainly triglycerides (9697%) followed by small quantity
of diglycerides (12%) and a variable quantity of free fatty
acids which are used as a marker of oil quality (Olias and
Garcia, 1997).
Fig. 1A shows the changes in the percent free fatty
acids (FFA) (% oleic acid) of the oils obtained from olives
stored at the dierent temperatures and under dierent
atmospheres. The oils extracted immediately post-harvest
had an average FFA value of 0.25% 0.04 and were
therefore categorized as extra virgin olive oil. During
the initial 15 days of storage, the FFA values of the oils
from all three treatments were lower than 0.8%, which
is the limit set for extra virgin olive oil quality in
ECC/1989/03. The oils obtained from the olives stored
at 5 C for 30 days had FFA values within the limit of
the extra virgin category irrespective of storage atmo-
sphere. By contrast, the oil extracted from the olives kept
at ambient temperature had an FFA value >2 and was
classied as lampante olive oil . Garcia, Gutierrez,
Castellano et al. (1996) reported that Picual olives stored
at 5 C for 45 days retained FFA value levels <1%, this
0
0 15 30
0 15 30
0 15 30 0 15 30
0 15 30
0 15 30
0.5
1
1.5
2
2.5
Storage time (days)
P
e
r
c
e
n
t

f
r
e
e

f
a
t
t
y

a
c
i
d
s

(
%

o
l
e
i
c

a
c
i
d
)
0.00
2.00
4.00
6.00
8.00
Storage time (days)
P
e
r
o
x
i
d
e

v
a
l
u
e

(
m
e
q
/
k
g
)
1.50
1.80
2.10
2.40
Storage time (days)
K
2
3
2
0.00
0.10
0.20
0.30
0.40
0.50
Storage time (days)
K
2
7
0
250
300
350
400
450
Storage time (days)
T
o
t
a
l

p
h
e
n
o
l
s

(
p
p
m
)


7
9
11
13
15
17
19
Storage time (days)
I
n
d
u
c
t
i
o
n

t
i
m
e

(
h
;

1
2
0

C
;

2
0
l

O
2
/
h
)
A B
D C
E F
Fig. 1. Changes in the percent free fatty acids (%oleic acid) (A), in the peroxide value (meq/kg) (B), in the content of conjugated fatty acids measured by the
specic extinction coecient at 232 nm(K232) (C), in the content on carbonylic compounds measured by the specic extinction coecient at 270 nm(K270)
(D), in the total phenol content (ppm) (E) and in the stability to oxidation (hours) (F) of oils obtained fromolives Coratina stored at dierent temperatures
and atmospheres. Each data point represents the mean of two replicates (d, 3%O
2
+ 5%CO
2
at 5 C; j, humidied air 5 C; m, ambient 20 C).
M.L. Clodoveo et al. / Food Chemistry 102 (2007) 571576 573
result is similar to our observations with Coratina olives
The increase in the FFA value of an oil during storage
is positively related to increasing storage temperature
(Garcia et al., 1994; Gutierrez, Perdiguero, Garcia et al.,
1992). Gutierrez, Perdiguero, Garcia et al. (1992) reported
that the increase in the acidity of oils extracted from fruit
stored at 20 C correlated well with decay incidence. In
general, the rst action of a parasitic microorganism in
an oil-rich tissue is the to induce hydrolytic activity by lip-
ases which leads to the release of fatty acids from the tri-
acylglycerol molecules of the oil. The free fatty acids are
easily metabolized by microorganisms. Fruit storage at
5 C allowed the FFA of the oil to be maintained at
0.8% for up to 30 days.
3.2. Peroxide value
The peroxide value (PV) is a measure of primary oxida-
tion. Fig. 1B shows the changes in the PV (meq O
2
/kg) of
oils obtained from olives stored at the dierent tempera-
tures and under dierent atmospheres. The PV of the oils
extracted immediately after the harvesting was
2.0 meq O
2
/kg. Cold storage at 5 C signicantly delayed
the rise in the peroxide value of the oils. The PV of the
oil obtained from olives stored at 20 C for 15 days was
double that of the baseline values. At the end of observa-
tion trial (30 days), none of the oils analyzed exceeded
the maximum peroxide value for extra virgin olive oil
(20 meq O
2
/kg) since the highest PV obtained was on aver-
age 5.8 meq O
2
/kg for oils extracted from olives stored at
ambient temperature for 30 days. This result is due to the
high phenol content of the Coratina cultivar which pre-
serves the oil from oxidative phenomena.
3.3. Specic extinction coecient at 232 nm and 270 nm
The K
232
parameter is mainly indicative of the conjuga-
tion of trienes and also of the presence of carbonylic com-
pounds. The maximum permitted values of K
232
and K
270
for extra virgin olive oils are 2.50 and 0.20, respectively.
Fig. 1C and D show the changes in K
232
and K
270
of oils
obtained from olives stored at dierent temperatures and
under dierent atmospheres. The K
232
of the oils extracted
immediately post-harvest was on average 1.63 and the K
270
was 0.13. The K
232
did not change markedly in the oil from
olives stored at 5 C. The oil from olives stored at 20 C
had a higher K
232
value (2.27 after 30 days). A K
232
value
of 2.50, which is the limit for extra virgin olive oil, was
not exceeded irrespective of storage atmosphere and tem-
perature. Storage of olives at a low temperature had a posi-
tive eect on the K
270
of the oils; 0.22 (the limit for extra
virgin olive oil) was not surpassed in any of the treatments
with the exception of the oil obtained from olives stored at
20 C (on average 0.43 0.06). Our results for the K
270
val-
ues are similar to those of previous reports (Garcia, Gut-
ierrez, Castellano et al., 1996; Gutierrez, Perdiguero,
Garcia et al., 1992).
3.4. Total phenol content
Virgin olive oil contains phenolic compounds responsi-
ble for its fragrance and peculiar avour, (Gutierrez, Perd-
iguero, Gutierrez, and Olias, 1992; Servili et al., 2004).
These substances also contribute to the oxidative stability
of the oil (Baldioli, Servili, Perretti, and Montedoro,
1996; Gutnger, 1981; Papadopoulus and Boskow, 1991)
and protect consumers against cancer and atherosclerosis
(La Vecchia et al., 1995; Trichopoulou et al., 1995; Visioli,
Bellosta, and Galli, 1997). Fig. 1E shows the changes in the
total phenol content (mg of gallic acid/kg of oil) of the oils
obtained from olives stored at dierent temperatures and
under dierent atmospheres. The total phenols decreased
in the oils obtained form olives kept at the higher temper-
ature and in ambient air, where the concentration of the
oxygen was higher than the controlled atmosphere.
During storage uid exuding from the olives may pro-
vide an optimum medium for the growth of fungi and bac-
teria (Olias and Garcia, 1997). Pseudomonas and other soil
bacteria are able to metabolize a wide variety of aromatic
compounds, such as phenol and its derivatives (Watanabe,
Hino, Onodera, Kajie, and Takahashi, 1996). Low temper-
atures may have a bacteriostatic action that controls micro-
bial development. Moreover, breakdown of the cells may
favour contact of the phenolic substances with the oxida-
tive enzymes. Olives contain oxidoreductases, such as poly-
phenoloxidase and peroxidase, that may oxidize
polyphenols and impair the health-related qualities and
sensory characteristics of olive oil (Servili, Selvaggini, Tat-
icchi, Esposto, and Montedoro, 2003), the activity of these
enzymes is known to decrease at low temperatures. The
results we obtained are consistent with these observations.
3.5. Oxidative stability
The antioxidant activity of hydrophilic phenols of virgin
olive oil has been extensively studied (Servili et al., 2004).
As reportedbyseveral investigators, the concentrationof phe-
nolic compounds, evaluatedcolorimetrically andexpressedas
total phenols, shows a correlation with the shelf -life of virgin
olive oil as tested by accelerated methods such as Rancimat
(Gutierrez Gonzales-Quijano, Janer del Valle, Janer del Valle,
Gutierrez Rosales and Vazquez Roncero, 1977).
Fig. 1F shows the changes in oxidation stability the of
the oils obtained from olives stored at dierent tempera-
tures and under dierent atmospheres. Fig. 2 relates total
phenols to the corresponding induction times for the oils
obtained from the olives stored under the three conditions
described. The overall values show that a direct correlation
exists between the two parameters. The regression line
computed yielded r
2
= 0.88 (p < 0.05).
3.6. Polar compounds and triglyceride oligopolymers
The HPSEC method measured the triglyceride
oligopolymers (PTGs), oxidized triglycerides (ox-TGs)
574 M.L. Clodoveo et al. / Food Chemistry 102 (2007) 571576
and diglycerides (DGs). PTGs are absent or present only in
trace amounts in good quality olive oils. Ox-TGs include
all forms of oxidation of triglycerides and are an important
class of degradation substances so their determination
helps to better dene the oxidative degradation of an oil.
Fig. 3A shows the changes in the ox-TG (%) of oils
obtained from olives stored at dierent temperatures and
under dierent atmospheres. Determination of DGs in oils
is useful to quantify the hydrolytic degradation of an oil.
Fig. 3B shows the changes in the DGs (%) of oils obtained
from olives stored at dierent temperatures and under dif-
ferent atmospheres.
Table 1 shows the ANOVA analysis of oxidized triglyc-
erides of oils extracted from olives immediately post-har-
vest and of oils extracted from olives after 15 and 30 days
of storage at dierent temperatures and under dierent
atmospheres. The data represent the mean (M) (g/100 g
of oil) Standard deviation (SD). Duration of olive stor-
age signicantly aected the amount of ox-TG
(p < 0.01%) in all the experimental conditions considered.
The ox-TG values diered in a statistically signicant way
(p < 0.01) for the three conditions applied also over the
same duration of storage. At the end of the storage per-
iod, the lowest ox-TG was obtained when the drupes were
stored at 5 C in humidied air and at 5 C under con-
trolled atmosphere. When the drupes were stored in air
at 20 C the oils extracted contained much higher
amounts of ox-TG. An average 30% more was found than
in the oils obtained from olives stored at 5 C in humid-
ied air and an average 12% more was found than in
the oils obtained from olives stored at 5 C in controlled
atmosphere. When the percent values of ox-TG were plot-
ted against the corresponding induction times a linear
trend of the experimental data was depicted for all the
series of samples examined (Fig. 4). The gure shows that
a inverse correlation was found (p < 0.05%). The fact that
the amount of ox-TG is inversely correlated to the induc-
tion time is further support of the value of this parameter
in dening the oxidative degradation of oils and in pre-
dicting their shelf life.
7
250 300 350 400 450
9
11
13
15
17
19
Total phenols (ppm)
I
n
d
u
c
t
i
o
n

t
i
m
e

(
h

;

1
2
0

C
;

2
0
l

O
2
/
h
)
r = 0.9405
Fig. 2. Correlation between total phenols and induction time.
0.40
0.50
0.60
0.70
0.80
0.90
0 15 30 0 15 30
Storage time (days)
O
x
i
d
i
z
e
d

t
r
i
g
l
y
c
e
r
i
d
e
s


(
%
)
1.00
1.40
1.80
2.20
2.60
3.00
Storage time (days)
D
i
g
l
y
c
e
r
i
d
e
s

(
%
)
A B
Fig. 3. Changes in oxidized triglycerides (%) (A) and in diglycerides (%) (B) of oils obtained from olives Coratina stored at dierent temperatures and
atmospheres. Each data point represents the mean of two replicates (d, 3%O
2
+ 5%CO
2
at 5 C; j, humidied air 5 C; m, ambient 20 C).
Table 1
ANOVA analysis of oxidized triglycerides of oils extracted from olives immediately post-harvest and of oils extracted from olives after 15 and 30 days of
storage at dierent temperatures and under dierent atmospheres
Storage treatment 3%O
2
+ 5% CO
2
at 5 C Humidied air at 5 C Ambient 20 C
Days M SD M SD M SD
0 0.43 0.01aA 0.43 0.01aA 0.43 0.01aA
15 0.64 0.01aB 0.58 0.01bB 0.79 0.01cB
30 0.74 0.02aC 0.64 0.01bC 0.83 0.02cC
Data represents mean (M) (g/100 g of oil) and standard deviation (SD).
Signicant dierences in the same row are showed by dierent small letters (P < 0.01).
Signicant dierences in the same column are showed by dierent capital letters (P < 0.01).
M.L. Clodoveo et al. / Food Chemistry 102 (2007) 571576 575
4. Conclusions
This experimentation provided evidence that, after
thirty days, oils obtained from olives stored at 5 C pre-
served the best characteristics compared to those obtained
from olives kept at ambient temperature. Ensuring a con-
trolled atmosphere during storage did not produce any
remarkable advantage over storage in humidied air at
5 C. Therefore, considering the cost of controlled atmo-
sphere technology, the best option for storing Coratina
olives is to simply keep them at cold temperatures.
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7
0.4 0.5 0.6 0.7 0.8 0.9
9
11
13
15
17
19
Oxidized triglycerides (%)
I
n
d
u
c
t
i
o
n

t
i
m
e

(
h

a
t

1
2
0

C
;

2
0
l

O
2
/
h
)
r = 0.7803
Fig. 4. Correlation between oxidized triglycerides and Induction time.
576 M.L. Clodoveo et al. / Food Chemistry 102 (2007) 571576